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PMID: 8278360 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The frequency of gene targeting in yeast depends on the number of target copies.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 91 ·No. 1 ·1994-01-04 ·Pages 177-81

Wilson JH, Leung WY, Bosco G, Dieu D, Haber JE

Abstract

We have compared the efficiency of transformation by linear DNA fragments in yeast strains carrying different numbers of homologous targets for recombination. In strains carrying dispersed copies of a target and in strains carrying tandem arrays, the frequency of transformation is proportional to the number of targets. This result is in contrast to previous studies of transformation in mammalian cells, where targeted integration was insensitive to the number of targets. We conclude that, in yeast, the search for a homologous partner is a rate-limiting step in the successful recombination of linearized DNA fragments. Furthermore, the fact that we obtain the same results with both dispersed and clustered targets argues against models of homology searching in which DNA becomes nonspecifically associated with a chromosome and then slides along the DNA until homology is encountered.

MeSH Terms
Chromosome Mapping DNA, Fungal/genetics Genes, Fungal Recombination, Genetic Restriction Mapping Saccharomyces cerevisiae/genetics Transformation, Genetic
Chemicals
DNA, Fungal
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wilson J H
Department of Biochemistry, Baylor College of Medicine, Houston, TX 77030.
Leung W Y
Bosco G
Dieu D
Haber J E
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.91.1.177
Published
1994-01-04
Pages
177-81
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC42909
Subset
IM
Grants
NIGMS NIH HHS · GM29736 · United States
NIGMS NIH HHS · GM38219 · United States
External Links
PubMed source DOI: 10.1073/pnas.91.1.177 PMC Full Text View journal details
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